Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI32458
Trapped Gene
D230025D16Rik (ENSMUSG00000031889)
Vector Insertion
Chr 8: 107763973 - 107764740
Public Clones IST10890A7 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 36% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000236883 (Chr8:107763878..107763972 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GAGCAGCTCTTCCACCTCAA Chr8:107763892..107763911 60.68 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000236883 (Chr8:107763878..107763972 +)
Downstram Exon
ENSMUSE00000236858 (Chr8:107764741..107764838 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GAGCAGCTCTTCCACCTCAA Chr8:107763892..107763911 60.68 55 AAGCTAGACCATGGGCAAAA Chr8:107764768..107764787 59.71 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000405558 Chr8:107749099..107749386 CGAGCAATGGGAATTCACTC Chr8:107749364..107749383 60.6 50
upstream ENSMUSE00000236963 Chr8:107755028..107755110 AAGCACTGTCGCATCATCAG Chr8:107755063..107755082 60.02 50
upstream ENSMUSE00000236942 Chr8:107758382..107758465 GACTCAGGACGGGATCACAC Chr8:107758414..107758433 60.54 60
upstream ENSMUSE00000236921 Chr8:107762323..107762366 No primer for this exon
upstream ENSMUSE00000236900 Chr8:107763571..107763650 CCATCGCTCCTACCATTGAG Chr8:107763594..107763613 60.62 55
upstream ENSMUSE00000236883 Chr8:107763878..107763972 GAGCAGCTCTTCCACCTCAA Chr8:107763892..107763911 60.68 55

*** Putative Vector Insertion (Chr 8: 107763973 - 107764740) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000236858 Chr8:107764741..107764838 AAGCTAGACCATGGGCAAAA Chr8:107764768..107764787 59.71 45
downstream ENSMUSE00000236840 Chr8:107764927..107765039 GGTCCAGTTCCATCTCGAAG Chr8:107765010..107765029 59.65 55
downstream ENSMUSE00000214347 Chr8:107770321..107770454 CGCACGCTCAAATACTCTCA Chr8:107770376..107770395 60.16 50
downstream ENSMUSE00000214340 Chr8:107770738..107770821 GTTGCACTTGGACGGAACTT Chr8:107770791..107770810 60.16 50
downstream ENSMUSE00000214342 Chr8:107771996..107772078 TGCCCAGGGTAATTGGTATG Chr8:107772066..107772085 60.58 50
downstream ENSMUSE00000214344 Chr8:107772643..107772686 TGAACTCACAGCGGTGGTAA Chr8:107772665..107772684 60.3 50
downstream ENSMUSE00000214348 Chr8:107773319..107773362 CAGTCTGACCACCTGCATTTT Chr8:107773341..107773361 60.16 47.62
downstream ENSMUSE00000214346 Chr8:107773497..107773555 GCAAGACAACAGGCTTCTCC Chr8:107773554..107773573 60 55
downstream ENSMUSE00000214341 Chr8:107774619..107774691 GCCAAACGGATTGGTGTTAT Chr8:107774655..107774674 59.69 45
downstream ENSMUSE00000377433 Chr8:107775498..107776949 CTGTAGGCCCAGGAACCATA Chr8:107776741..107776760 59.95 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GCACTTCTGTGTCCTTGTGG Chr8:107763978..107763998 59.31 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GCAGGACACCTTTCGTGACT Chr8:107764011..107764031 60.31 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000031889